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Evaluating the Feasibility of Low Cost Sperm Preparation Methods Within a Prospective Intrauterine Insemination Programme in Gabon

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Evaluating the Feasibility of Low Cost Sperm Preparation Methods Within a Prospective Intrauterine Insemination Programme in Gabon Zzz Evaluating the feasibility of low cost sperm preparation methods within a prospective intrauterine insemination programme in Gabon By Lionel Wildy Moungala (12232310) Submitted in fulfilment of the requirements for the degree Masters of Science (Reproductive Biology) In the Faculty of Health Sciences University of Pretoria September 2015 Supervisor: Prof C. Huyser (PhD) Reproductive Biology Laboratory Department of Obstetrics and Gynaecology Reproductive Endocrine Unit University of Pretoria Steve Biko Academic Hospital i DEDICATION Every challenging work needs self-effort as well as guidance of elders, especially those who are very close to our heart. I dedicate my humble effort to my sweet and loving God, the Reproductive Biology Laboratory members, my mother (Constance Moungala) and my dad (William Moungala) whose affection, love, encouragement and prayers make me able to complete this journey i DECLARATION BY CANDIDATE „I hereby declare that the dissertation submitted for the degree MSc Reproductive Biology, in the Faculty of Health Sciences, University of Pretoria is my own original work and has not previously been submitted to any other institution of higher education. I further declare that all sources cited or quoted are indicated and acknowledged by means of a comprehensive list of references.‟ LIONEL WILDY MOUNGALA Name of student Signature Date ii ETHICAL CLEARANCES iii iv Contents DEDICATION..................................................................................................................................... i DECLARATION BY CANDIDATE ........................................................................................................ii ETHICAL CLEARANCES .................................................................................................................... iii ACKNOWLEDGEMENTS .................................................................................................................. 9 SUMMARY .................................................................................................................................... 10 LIST OF ABBREVIATIONS .............................................................................................................. 12 LIST OF FIGURES ........................................................................................................................... 13 LIST OF TABLES ............................................................................................................................. 15 CHAPTER 1: OVERVIEW OF STUDY........................................................................................................ 16 1.1 Motivation for study ................................................................................................................. 16 1.2 Research questions ................................................................................................................... 19 1.3 Hypotheses ................................................................................................................................ 19 1.4 Objectives .................................................................................................................................. 20 CHAPTER 2: LITERATURE REVIEW ......................................................................................................... 21 2.1 Introduction .................................................................................................................................... 21 2.2 Overview of infertility in Africa ...................................................................................................... 22 2.2.1 Prevalence of childlessness .............................................................................................. 22 2.2.2 Impact of infertility .......................................................................................................... 28 2.3 Factors associated with infertility .................................................................................................. 30 2.3.1 Anatomical and pathological causes of infertility ............................................................ 30 2.3.2 Sexually transmitted infections and Human immunodeficiency virus ............................ 33 2.3.3 Behavioural and lifestyle factors ...................................................................................... 34 2.4 Sperm preparation methods .......................................................................................................... 37 2.4.1 Standard swim-up method .............................................................................................. 38 5 2.4.2 SEP-D kit . …………………………………………………………………………………………………………………….39 2.4.3 Single layer centrifugation (SLC) ...................................................................................... 40 2.4.4 The density gradient centrifugation (DGC) ...................................................................... 40 2.5 Assisted Reproductive Technology (ART) treatments .................................................................... 41 2.5.1 Intrauterine insemination (IUI) ........................................................................................ 41 2.5.3 In vitro fertilization (IVF) .................................................................................................. 43 2.5.4 Intra-cytoplasmic sperm injection (ICSI) .......................................................................... 44 2.6 Access to infertility treatment in Africa ......................................................................................... 44 2.7 Conclusion ...................................................................................................................................... 46 CHAPTER 3: MATERIALS AND METHODS ............................................................................................. 47 3.1 Background ..................................................................................................................................... 47 3.2 Sampling and semen collection ...................................................................................................... 47 3.3 Experimental set-up ....................................................................................................................... 48 3.3.1 Experiment (i): Development of a simplified sperm swim-up method to obtain motile sperm with minimum equipment ............................................................................................. 48 3.3.2 Experiment (ii): Comparison between the simplified sperm swim-up (SW-10) method and the commercially available SEP-D kit ................................................................................. 50 3.3.3 Experiment (iii): Comparison between the single layer and density gradient centrifugation, based on post-processed sperm parameters .................................................. 52 3.4 Sperm analyses ............................................................................................................................... 53 3.4.1 Motility (Total motile count): Experiment (i), (ii) and (iii) ................................................ 53 3.4.2 Sperm vitality: Experiment (ii) ......................................................................................... 55 3.4.3 DNA fragmentation .......................................................................................................... 57 3.4.4 Sperm morphology .......................................................................................................... 58 3.5 Evaluation of the feasibility of establishing an IUI programme in Libreville (Gabon) .................... 59 3.6 Statistical analyses .......................................................................................................................... 60 3.6.1 Experiment (i), (ii) and (iii) ............................................................................................... 60 3.6.2 Feasibility of establishing an IUI programme in Libreville (Gabon) ................................. 60 6 CHAPTER 4: RESULTS ............................................................................................................................... 61 4.1 Introduction .................................................................................................................................... 61 4.2 Section 1: Development of a simplified swim-up method ............................................................. 62 4.2.1 Experiment (i): Total motile count of spermatozoa obtained after processing using 5 ml (SW-5), 10 ml (SW-10) and 20 ml (SW-20) syringes .......................................................... 62 4.2.2 Experiment (ii): Sperm parameters obtained from the simplified sperm swim-up (SW-10) method and the commercial SEP-D kit ....................................................................... 66 4.2.3 Experiment (iii): Concentration and motility parameters of spermatozoa after processing using single layer centrifugation (SLC) and double layer density gradient centrifugation (DGC) ................................................................................................................. 69 4.3 Section 2: Gynaecologists’ opinions on the diagnosis and treatment of infertility in Gabon ........ 72 4.3.1 Gynaecologists’ access to abdominal/vaginal ultrasound scans and an estimate of infertility-related consultations ...............................................................................................
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